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NF-kB p65 抗体

Name: NF-kB p65 抗体
Brand: antibodies-online
SKU: ABIN6135887
Availability: OutOfStock
Rating: 5 (1 reviews)

NFkBP65 适用: 人 WB, IHC, IP, IF 宿主: 小鼠 Monoclonal unconjugated

Immunofluorescence validation image for anti-Nuclear Factor-kB p65 (NFkBP65) antibody (ABIN6135887) (NF-kB p65 抗体)

Western blot analysis of extracts of various cell lines, using RELA antibody. (NF-kB p65 抗体)

2 images

(1 reference)

(1 validation)

产品编号 ABIN6135887

发货至: 中国

Datasheet as PDF

抗原 See all NF-kB p65 (NFkBP65) 抗体

NF-kB p65 (NFkBP65) (Nuclear Factor-kB p65 (NFkBP65))
适用
490

348

281

40

34

28

23

19

19

14

14

10

5

2

1

1

1
人
宿主
450

42

3

2

1
小鼠
Compatible Secondaries
克隆类型
432

66
单克隆
标记
262

25

19

14

10

10

10

10

10

10

10

10

10

10

10

10

9

8

8

8

8

8

5

2

2
This NF-kB p65 antibody is un-conjugated
应用范围
390

177

130

119

104

100

85

70

70

28

25

9

7

7

7

6

4

3

2

2

1
Western Blotting (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), Immunofluorescence (IF)

交叉反应

人, 小鼠, 大鼠

产品特性

Monoclonal Antibodies

免疫原

Recombinant protein of human NF-kB p65

亚型

IgG

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anti-Nuclear Factor-kB p65 (NFkBP65) (Center) antibody
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anti-Nuclear Factor-kB p65 (NFkBP65) (AA 19-306) antibody
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anti-Nuclear Factor-kB p65 (NFkBP65) (acLys314), (acLys315) antibody
NFkBP65 适用: 人, 小鼠, 大鼠 WB, ELISA, IHC (p) 宿主: 兔 Polyclonal unconjugated

anti-Nuclear Factor-kB p65 (NFkBP65) (AA 50-180) antibody
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anti-Nuclear Factor-kB p65 (NFkBP65) (pThr435) antibody
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anti-Nuclear Factor-kB p65 (NFkBP65) (pSer281) antibody
NFkBP65 适用: 人, 小鼠, 大鼠 WB, IHC, ELISA, IF, ICC 宿主: 兔 Polyclonal unconjugated

应用备注

WB,1:500 - 1:2000,IHC,1:50 - 1:200,IF,1:50 - 1:100,IP,1:20 - 1:50

说明

HIGH QUALITY

限制

仅限研究用
生效 #104075 (Immunofluorescence)

by

Klinik für Anästhesiologie, Intensivmedizin und Schmerztherapie des Universitätsklinikums Knappschaftskrankenhaus Bochum

No.

#104075

日期

2019.11.21

抗原

NfkBP65

Lot Number

9410609001

Method validated

Immunofluorescence

Positive Control

U-937 human monocytic cell line

Negative Control

No primary antibody control

Notes

Passed. ABIN6135887 specifically stains RelA/p65 in U-937 human monocytic cells with antigen retrieval using 1% SDS.

Validation Images

U937 monocytic cells stained with with anti- RelA/p65 antibody ABIN6135887 and an AF488-labeled secondary antibody (bottom row, green). Counterstain with DAPI (blue). Negative control without primary antibody (top row) shows no AF488 signal.
Full Methods
Primary Antibody

ABIN6135887

Secondary Antibody

AF488-conjugated AffiniPure alpaca anti-mouse IgG antibody (Jackson Immuno Research, 615-545-214)

Full Protocol

Grow U-937 cells in RPMI 1640 Medium (Gibco, 21875034) supplemented with 10 % FCS (Gibco, A3840001) and Pen/Strep (Gibco, 15140122), at 37°C and 5% CO₂ in 300µL in 24-well plates (Sarstedt AG&Co. KG).
Use cytospin to spin cells onto slide.
Fix cells on coverslide in 4% paraformaldehyde (Carl Roth GmbH, 0335.1) for 30min at RT.
Wash cells 3x for 5min with PBS (Sigma-Aldrich, 8537).
Permeabilize cells in 100µL PBS containing 0.1 % Triton X-100 (Sigma-Aldrich, T8787) for 5min at RT.
Wash cells 3x for 5min with PBS.
Permeablilize cells in 100 µL PBS containing 1 % SDS (0183, Carl Roth GmbH) for 5 min at RT.
Wash cells 3x for 5 min with PBS.
Block non-specific binding with 1x Duolink Blocking solution (Sigma-Aldrich, DUO82007) for 30min at RT.
Incubate cells with primary mouse anti-human RelA/p65 antibody (antibodies-online, ABIN6135887, lot 9410609001) diluted 1:100 in 1x Duolink Antibody diluent (Sigma-Aldrich, DUO82008) ON at 4°C.
Wash cells 3x for 5min with PBS.
Incubate cells with secondary AF488-conjugated AffiniPure alpaca anti-mouse IgG antibody (Jackson Immuno Research, 615-545-214) diluted 1:400 in 1x Duolink Antibody diluent for 1h at RT in the dark.
Wash cells 2x for 5min with PBS.
Mount coverslips on glass slides in 50µL ProLong Gold antifade mountant with DAPI (Invitrogen, P36931).
Image acquisition with an Olympus widefield microscope.

Experimental Notes

Strong green staining dependent on the presence of primary antibody indicates that the antibody works. Negative control lacking the primary antibody shows only background fluorescence. Staining was obtained using antigen retrieval (1% SDS). Without this the antibody did not yield detectable signal.
状态

Liquid

缓冲液

PBS with 0.02 % sodium azide,50 % glycerol, pH 7.3.

储存液

Sodium azide

注意事项

This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

储存条件

-20 °C

储存方法

Store at -20°C. Avoid freeze / thaw cycles.
Fang, Fang, Ren, Hong, Liu, Zhao, Wang, Peng, Xiao: "Porcine Deltacoronavirus Accessory Protein NS6 Antagonizes Interferon Beta Production by Interfering with the Binding of RIG-I/MDA5 to Double-Stranded RNA." in: Journal of virology, Vol. 92, Issue 15, (2018) (PubMed).
抗原

NF-kB p65 (NFkBP65) (Nuclear Factor-kB p65 (NFkBP65))

别名

RELA (NFkBP65 产品)

背景

NF-kappa-B is a ubiquitous transcription factor involved in several biological processes. It is held in the cytoplasm in an inactive state by specific inhibitors. Upon degradation of the inhibitor, NF-kappa-B moves to the nucleus and activates transcription of specific genes. NF-kappa-B is composed of NFKB1 or NFKB2 bound to either REL, RELA, or RELB. The most abundant form of NF-kappa-B is NFKB1 complexed with the product of this gene, RELA. Four transcript variants encoding different isoforms have been found for this gene.,NFKB3,p65,NF-kB p65,RELA,CMCU,Epigenetics & Nuclear Signaling,Transcription Factors,Cancer,Signal Transduction,MAPK-P38 Signaling Pathway,Cell Biology & Developmental Biology,Apoptosis,Inhibition of Apoptosis,Death Receptor Signaling Pathway,Endocrine & Metabolism,Endocrine and metabolic diseases,Obesity,Immunology & Inflammation,B Cell Receptor Signaling Pathway,T Cell Receptor Signaling Pathway,IL-6 Receptor Signaling Pathway,NF-kB Signaling Pathway,Toll-like Receptor Signaling Pathway,Neuroscience,Neurodegenerative Diseases,Amyloid Plaque and Neurofibrillary Tangle Formation in Alzheimer's Disease,Cardiovascular,RELA

分子量

58 kDa/59 kDa/60 kDa

基因ID

5970

UniProt

Q04206

途径

NF-kappaB Signaling, RTK signaling, TCR Signaling, TLR signaling, Fc-epsilon Receptor Signaling Pathway, Neurotrophin Signaling Pathway, Activation of Innate immune Response, Cellular Response to Molecule of Bacterial Origin, Hepatitis C, Toll-Like Receptors Cascades, S100 Proteins